Sample preparation optimization for the simultaneous determination of mycotoxins in cereals

Abstract

The efficiency of three extraction solvents and three clean-up procedures was compared for simultaneous extraction and purification of aflatoxins (AFB1, AFB2, AFG1 and AFG2), ochratoxin A (OTA), and zearalenone (ZEA) from spiked cereal samples. The best recovery rates for all mycotoxins were achieved using methanol: water (80:20) as the extraction solvent and AOZ multi-functional immunoaffinity column (IAC), as clean up method with recovery values of 61–89%, while that of Oasis HLB and MycoSep 226 were 37–67% and 44–78%, respectively. Then, five variables in the IAC clean-up conditions, including primary conditioning with phosphate buffer saline (PBS) (0–10 mL) (X1), extract load up volume (10–20 mL) (X2), washing volume with PBS (10–20 mL) (X3), and eluting solution volumes with methanol (1–3 mL) (X4) and acetic acid (0–1.5 mL) (X5), were optimized for the specific purification and enrichment of the mycotoxins. Results showed that primary conditioning and PBS washing did not have a significant effect on the recovery responses of mycotoxins. Optimized conditions were selected as 0, 15, 10, 1.3, and 1.5 mL for X1–X5, respectively. The recovery rates of AFB1, AFB2, AFG1, AFG2, OTA and ZEA were within 93–104% in spiked rice, under optimal conditions. LOD and LOQ were 0.0125 and 0.05 ng/g for AFB1 and AFG1, 0.0037 and 0.015 ng/g for AFB2 and AFG2, 0.05 and 0.2 ng/g for OTA, and 0.5 and 2 ng/g for ZEA, respectively. Extraction of spiked cereal samples with methanol: water (80:20) and clean up using AOZ IAC column in optimal condition provided recovery range of 77–104% for all targeted mycotoxins.

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References

  1. 1.

    Göbel R, Lusky K (2004) J AOAC Int 87:411

    Google Scholar 

  2. 2.

    WHO (2001) In: W. H. Organization (ed) Food additives series, vol 47, Food Additives Series. WHO, Geneva

  3. 3.

    Trucksess M, Weaver C, Oles C, D’Ovidio K, Rader J (2006) J AOAC Int 89:624

    CAS  Google Scholar 

  4. 4.

    Stecher G, Jarukamjorn K, Zaborski P, Bakry R, Huck CW, Bonn GK (2007) Talanta 73:251

    Article  CAS  Google Scholar 

  5. 5.

    EC/466/2001. In: Official Journal of the European Union, vol L 77/1, Official Journal of the European Union, pp 6

  6. 6.

    EC/472/2002, Vol L 75/18. Official Journal of the European Union, pp 1

  7. 7.

    EC/683/2004, Vol L 106/3. Official Journal of the European Union, pp 1

  8. 8.

    EC/856/2005, Vol L 143/3. Official Journal of the European Union, pp 1

  9. 9.

    EC/1881/2006, Vol L 364/5. Official Journal of the European Union, pp 15

  10. 10.

    Krska R, Josephs R (2001) Fresenius J Anal Chem 369:469

    Article  CAS  Google Scholar 

  11. 11.

    Trucksess MW, Weaver CM, Oles CJ, Fry FS Jr, Noonan GO, Betz JM, Rader JI (2008) J AOAC Int 91:511

    CAS  Google Scholar 

  12. 12.

    Danicke S, Ueberschar KH, Valenta H, Matthes S, Matthaus K, Halle I (2004) Br Poult Sci 45:264

    Article  CAS  Google Scholar 

  13. 13.

    Ofitserova M, Nerkar S, Pickering M, Torma L, Thiex N (2009) J AOAC Int 92:15

    CAS  Google Scholar 

  14. 14.

    Berthiller F, Werner U, Sulyok M, Krska R, Hauser MT, Schuhmacher R (2006) Food Addit Contam 23:1194

    Article  CAS  Google Scholar 

  15. 15.

    Ventura M, Guillen D, Anaya I, Broto-Puig F, Lliberia JL, Agut M, Comellas L (2006) Rapid Commun Mass Spectrom 20:3199

    Article  CAS  Google Scholar 

  16. 16.

    Berthiller F, Schuhmacher R, Buttinger G, Krska R (2005) J Chromatogr A 1062:209

    Article  CAS  Google Scholar 

  17. 17.

    Zollner P, Mayer-Helm B (2006) J Chromatogr A 1136:123

    Article  Google Scholar 

  18. 18.

    Lattanzio VM, Solfrizzo M, Powers S, Visconti A (2007) Rapid Commun Mass Spectrom 21:3253

    Article  CAS  Google Scholar 

  19. 19.

    Rahmani A, Jinap S, Soleimany F (2009) Compr Rev Food Sci Food Safety 8:202

    Article  CAS  Google Scholar 

  20. 20.

    Qiao J, Qi L, Ma H, Liu H, Yang J, Chen Y, Yang G (2009) Talanta 80:770

    Article  CAS  Google Scholar 

  21. 21.

    Hu YY, Zheng P, Zhang ZX, He YZ (2006) J Agric Food Chem 54:4126

    Article  CAS  Google Scholar 

  22. 22.

    Hennion MC, Pichon V (2003) J Chromatogr A 1000:29

    Article  CAS  Google Scholar 

  23. 23.

    Muronetz VI, Korpela T (2003) J Chromatogr B Analyt Technol Biomed Life Sci 790:53

    Article  CAS  Google Scholar 

  24. 24.

    Wang Y, Chai T, Lu G, Quan C, Duan H, Yao M, Zucker BA, Schlenker G (2008) Environ Res 107:139

    Article  CAS  Google Scholar 

  25. 25.

    Brum DM, Cassella RJ, Pereira Netto AD (2008) Talanta 74:1392

    Article  CAS  Google Scholar 

  26. 26.

    Myers RH, Montgomery DC, Anderson-cook CM (2009) Response surface methodology: process and product optimization using designed experiment, Wiley-Interscience Publication, NY, USA

    Google Scholar 

  27. 27.

    Sánchez-Romeu J, País-Chanfrau JM, Pestana-Vila Y, López-Larraburo I, Masso-Rodríguez Y, Linares-Domínguez M, Márquez-Perera G (2008) Biochem Eng J 38:1

    Article  Google Scholar 

  28. 28.

    Saez JM, Medina A, Gimeno-Adelantado JV, Mateo R, Jimenez M (2004) J Chromatogr A 1029:125

    Article  CAS  Google Scholar 

  29. 29.

    Ren Y, Zhang Y, Shao S, Cai Z, Feng L, Pan H, Wang Z (2007) J Chromatogr A 1143:48

    Article  CAS  Google Scholar 

  30. 30.

    EC/657/2002, Vol L221/8. Official Journal of the European Union, pp 1

  31. 31.

    Gilbert J, Anklam E (2002) TrAC Trends Anal Chem 21:468

    Article  CAS  Google Scholar 

  32. 32.

    EC/401/2006, Vol L70/12. Official Journal of the European Union, pp 31

  33. 33.

    Stroka J, Petz M, Joerissen U, Anklam E (1999) Food Addit Contam 16:331

    Article  CAS  Google Scholar 

  34. 34.

    Monbaliu S, Van Poucke C, Detavernier C, Dumoulin F, Van De Velde M, Schoeters E, Van Dyck S, Averkieva O, Van Peteghem C, De Saeger S (2010) J Agric Food Chem 58:66

    Article  CAS  Google Scholar 

  35. 35.

    Garon D, Richard E, Sage L, Bouchart V, Pottier D, Lebailly P (2006) J Agric Food Chem 54:3479

    Article  CAS  Google Scholar 

  36. 36.

    Richard E, Heutte N, Sage L, Pottier D, Bouchart V, Lebailly P, Garon D (2007) Food Chem Toxicol 45:2420

    Article  CAS  Google Scholar 

Download references

Acknowledgments

Authors acknowledge Universiti Putra Malaysia for financial support through Research University Grant Scheme (RUGS), Project Number 02-01-07-0024RU.

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Correspondence to S. Jinap.

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Rahmani, A., Jinap, S., Soleimany, F. et al. Sample preparation optimization for the simultaneous determination of mycotoxins in cereals. Eur Food Res Technol 232, 723–735 (2011). https://doi.org/10.1007/s00217-011-1431-y

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Keywords

  • Sample preparation
  • Optimization
  • Mycotoxin
  • Cereal
  • HPLC